Monstein Hans-Jurg, Grahn Niclas, Truedsson Mikael, Ohlsson Bodil
Molecular Biology Laboratory, Clinical Microbiology, University Hospital, Linkoping, Sweden.
World J Gastroenterol. 2006 Apr 28;12(16):2574-8. doi: 10.3748/wjg.v12.i16.2574.
To investigate the expression of gastrin-releasing peptide (GRP) and GRP-receptor mRNA in non-tumor tissues of the human esophagus, gastrointestinal tract, pancreas and gallbladder using molecular biology techniques.
Poly A(+) mRNA was isolated from total RNA extracts using an automated nucleic acid extractor and, subsequently, converted into single-stranded cDNA (ss-cDNA). PCR amplifications were carried out using gene-specific GRP and GRP-receptor primers. The specificity of the PCR amplicons was further confirmed by Southern blot analyses using gene-specific GRP and GRP-receptor hybridization probes.
Expression of GRP and GRP-receptor mRNA was detected at various levels in nearly all segments of the non-tumor specimens analysed, except the gallbladder. In most of the biopsy specimens, co-expression of both GRP and GRP-receptor mRNA appeared to take place. However, expression of GRP mRNA was more prominent than was GRP-receptor mRNA.
GRP and GRP-receptor mRNAs are expressed throughout the gastrointestinal tract and provides information for the future mapping and determination of its physiological importance in normal and tumor cells.
运用分子生物学技术,研究胃泌素释放肽(GRP)及GRP受体mRNA在人食管、胃肠道、胰腺和胆囊非肿瘤组织中的表达情况。
使用自动核酸提取仪从总RNA提取物中分离出Poly A(+) mRNA,随后将其转化为单链cDNA(ss-cDNA)。采用基因特异性GRP和GRP受体引物进行PCR扩增。使用基因特异性GRP和GRP受体杂交探针通过Southern印迹分析进一步确认PCR扩增产物的特异性。
在所分析的非肿瘤标本的几乎所有节段中,除胆囊外,均检测到不同水平的GRP和GRP受体mRNA表达。在大多数活检标本中,GRP和GRP受体mRNA似乎共同表达。然而,GRP mRNA的表达比GRP受体mRNA更显著。
GRP和GRP受体mRNA在整个胃肠道均有表达,为未来绘制其在正常细胞和肿瘤细胞中的分布图及其生理重要性的测定提供了信息。